How to amplify 1kb PCR product

Dr. Duncan Clark duncan at genesys.demon.co.uk
Mon Mar 2 12:38:14 EST 1998


In article <34FAD3B3.4B82 at public1.guangzhou.gd.cn>, gzzhjr <gzzhjr at PUBLI
C1.GUANGZHOU.GD.CN> writes
>Hello everyone,
>
>The length of DNA fragment I want to amplified by PCR is about 1kb,but I
>have no experience with this,could someone tell me the appropriate
>temperature and time in denature,annealing and expansion process?
>
>Thanks for any information.


For Taq polymerase use the usual extension time allowed is 1kb per
minute used at 72C. Denaturation on a block machine is usually 94C for
30 sec, annealing is for 30 secs at a temperature decided by your oligo
design. Use 2.5u Taq per 100ul PCR. 

Duncan
-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
TEl/FAX 01252376288
http://www.dnamp.com
http://www.genesys.demon.co.uk



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